Isolation of hydrogen generating microflora from cow dung for seeding anaerobic digester

The effectiveness of using cow dung as a source for isolating hydrogen generating microflora was investigated under varying isolating conditions based on viz.: pH adjustment and pH adjustment coupled with heat treatment. The viability of the isolated microflora was tested in an anaerobic jar with respect to biogas generation, hydrogen content and pH. The results showed that for pH adjusted microflora isolated from cow dung with solids content at 10% resulted in a cumulative biogas generation of 1494, 2404 and 3327 ml, whereas the corresponding cumulative hydrogen generation was found to be 424, 701 and 47 ml during the anaerobic fermentation for 120 h at a pH of 4, 5 and 6, respectively. The biogas was free from methane when operated at pH 4 and 5, whereas at pH 6 methane generation was observed. In the case of microflora isolated from cow dung with 10% solids, by subjecting to pH adjustment coupled with heat treatment resulted in biogas free from methane content during the fermentation at pH 4, 5 and 6, respectively. At the end of 120 h of fermentation for a reactor pH at 4, 5 and 6 the cumulative biogas generation was 1685, 2610 and 2353 ml, whereas the cumulative hydrogen generation was 509, 1198 and 1165 ml, respectively. A maximum of 41% and 62% hydrogen was obtained at pH 5 for microflora isolated based on pH adjustment and pH adjustment coupled with heat treatment. The effect of initial solids content of the cow dung on the isolating efficiency of hydrogen generating microflora was also investigated at pH 5 and 6 coupled with heat treatment. The results revealed that with the increase in initial solids content of the cow dung the optimum heat treatment period also increased as the pH increased from 5 to 6.     

Biohydrogen generation from anaerobic digestion of food waste

Biohydrogen generated from the anaerobic digestion of a synthetic food waste with constant composition and a real food waste collected in Hong Kong were studied. This study aims at using a monoculture to increase biohydrogen production and determining optimum conditions for maximum biohydrogen production. Among the nine bacteria screened for biohydrogen production, Escherichia cloacae and Enterobacter aerogenes produced the largest amount of biohydrogen from the anaerobic digestion of synthetic food waste. The optimum anaerobic digestion conditions were determined: initial pH of 7, a water to solids ratio of 5 (w/w), a mesophilic temperature (37 ± 1 °C), and in the presence of 40 mg/L FeSO₄·7H₂O. Anaerobic digestion at the optimum operating conditions using collected food waste with E. cloacae as the bacterial source was also performed. By adjusting the pH in the range of 5–6, a specific biohydrogen production of 155.2 mL/g of volatile solids (VS) in food waste was obtained.     

Biohydrogen production from poplar leaves pretreated by different methods using anaerobic mixed bacteria

Leaves are one of the main by-products of forestry. In this study, batch experiments were carried out to convert poplar leaves pretreated by different methods into hydrogen using anaerobic mixed bacteria at 35 °C. The effects of acid (HCl), alkaline (NaOH) and enzymatic (Viscozyme L, a mixture of arabanase, cellulase, β-glucanase, hemicellulase and xylanase) pretreatments on the saccharification of poplar leaves were studied. Furthermore, the effects of acid and enzymatic pretreatment on hydrogen production, together with their corresponding degradation efficiencies for the total reducing sugar (TRS) and metabolites were compared. A maximum cumulative hydrogen yield of 44.92 mL/g-dry poplar leaves was achieved from substrate pretreated with 2% Vicozyme L, which was approximately 3-fold greater than that in raw substrate and 1.34-fold greater than that from substrate pretreated with 4% HCl. The results show that enzymatic pretreatment is an effective method for enhancing the hydrogen yield from poplar leaves.     

Biohydrogen production from dual digestion pretreatment of poultry slaughterhouse sludge by anaerobic self-fermentation

Poultry slaughterhouse sludge from chicken processing wastewater treatment plant was tested for their suitability as a substrate and inoculum source for fermentation hydrogen production. Dual digestion of poultry slaughterhouse sludge was employed to produce hydrogen by batch anaerobic self-fermentation without any extra-seeds. The sludge (5% TS) was dual digested by aerobic thermophilic digestion at 55 °C with the varying retention time before using as substrate in anaerobic self-fermentation. The best digestion time for enriching hydrogen-producing seeds was 48 h as it completely repressed methanogenic activity and gave the maximum hydrogen yield of 136.9 mL H₂/g TS with a hydrogen production rate of 2.56 mL H₂/L/h. The hydrogen production of treated sludge at 48 h (136.9 mL H₂/g TS) was 15 times higher than that of the raw sludge (8.83 mL H₂/g TS). With this fermentation process, tCOD value in the activated sludge could be reduced up to 30%.     

菠萝皮厌氧发酵产沼气的研究

介绍了以菠萝皮为原料,在25℃恒温条件下,采用批量发酵工艺,进行发酵产沼气实验,实验结果表明,菠萝皮可以作为沼气发酵原料,未加碱时产沼气潜力为574.81 mL/gTS,604.30mL/gVS,甲烷含量51.34%;而加碱时产沼气潜力为568.66 mL/gTS,597.84 mL/gVS,甲烷含量52.96%。     

Biohydrogen production from cheese processing wastewater by anaerobic fermentation using mixed microbial communities

Hydrogen (H₂) production from simulated cheese processing wastewater via anaerobic fermentation was conducted using mixed microbial communities under mesophilic conditions. In batch H₂ fermentation experiments H₂ yields of 8 and 10 mM/g COD fed were achieved at food-to-microorganism (F/M)$(F/M)$ ratios of 1.0 and 1.5, respectively. Butyric, acetic, propionic, and valeric acids were the major volatile fatty acids (VFA) produced in the fermentation process. Continuous H₂ fermentation experiments were also performed using a completely mixed reactor (CSTR). The pH of the bioreactor was controlled in a range of 4.0–5.0 by addition of carbonate in the feed material. Maximum H₂ yields were between 1.8 and 2.3 mM/g COD fed for the loading rates (LRs) tested with a hydraulic retention time (HRT) of 24 h. Occasionally CH₄ was produced in the biogas with concurrent reductions in H₂ production; however, continuous H₂ production was achieved for over 3 weeks at each LR. The 16S rDNA analysis of DNA extracted from the bioreactors during periods of high H₂ production revealed that more than 50% of the bacteria present were members of the genus Lactobacillus and about 5% were Clostridia. When H₂ production in the bioreactors decreased concurrent reductions in the genus Lactobacillus were also observed. Therefore, the microbial populations in the bioreactors were closely related to the conditions and performance of the bioreactors.     

Biohydrogen production from specified risk materials co-digested with cattle manure

Biohydrogen production from the anaerobic digestion of specified risk materials (SRM) co-digested with cattle manure was assessed in a 3 × 5 factorial design. Total organic loading rates (OLR) of 10, 20, and 40 g L⁻¹ volatile solids (VS) were tested using manure:SRM (wt/wt) mixtures of 100:0 (control), 90:10, 80:20, 60:40, and 50:50 using five 2 L continuously stirred biodigesters operating at 55 °C. Gas samples were taken daily to determine hydrogen production, and slurry samples were analyzed daily for volatile fatty acid (VFA) concentration, total ammonia nitrogen (TAN), and VS degradation. Hydrogen production (mL g⁻¹ VS fed) varied quadratically according to OLR (P < 0.01), with maximum production at OLR20, while production decreased linearly (P < 0.0001) as SRM concentration increased. Reduced hydrogen production associated with SRM inclusion at >10% VS may be attributed to a rapid increase in TAN (r = −0.55) or other inhibitors such as long chain fatty acids. Reduced hydrogen production (P < 0.01) at OLR40 versus OLR20 may be related to increased rate of VFA accumulation and final VFA concentration (P < 0.001), as well as inhibition due to hydrogen accumulation (P < 0.001). Biohydrogen production from SRM co-digested with cattle manure may not be feasible on an industrial scale due to reduced hydrogen production with increasing levels of SRM.     

Production of hydrogen and methane from wastewater sludge using anaerobic fermentation

Hydrogen and methane were produced from wastewater sludge using a clostridium strain. The original sludge and pre-treated (acidified, basified and freeze/thawed) sludges were the testing samples. Some pre-treatments enhanced hydrogen yield, whereas other treatments enhanced methane yield. Hydrogen fermentation can be used as a pre-stage to improve subsequent methane production from wastewater sludge.     

An experimental approach to produce hydrogen and methane from food waste using catalyst

Anaerobic co-digestion of food waste, cow dung, and sludge solution is experimented in the presence of calcium peroxide (CaO₂) as the catalyst to produce hydrogen and methane as a source of renewable energy. The substrate to inoculum ratios (v/v) of 1:1(S1), 1:2(S2), 1:3(S3), 1:4(S4) and 1:5(S5) are investigated in separate fermentative and methanogenic reactors. The result from the fermentative reactors indicate maximum hydrogen concentration of 26.34% with cumulative yield of 114.1 mL/g total solid (TS) in S3 compared to the other samples. Methanogenic reaction shows the highest methane concentration of 54.13% in S3. The highest daily (average) and cumulative biogas yield of 5.36 mL/g TS and 201.9 mL/g TS respectively are identified in S3. A maximum carbon dioxide concentration of 63.11% is found in S1. Overall, the substrate to inoculum ratio of 1:3 is spotted to be optimal for effective hydrogen and methane production during the anaerobic co-digestion process.     

Biohydrogen production from mixed xylose/arabinose at thermophilic temperature by anaerobic mixed cultures in elephant dung

Biohydrogen production by batch fermentation of mixed xylose/arabinose at thermophilic temperature using anaerobic mixed cultures in elephant dung as the seed inoculums was investigated. Elephant dung was heat-treated in boiling water for 2 h before used as the seed inoculum in order to inhibit methanogenic activity. Biohydrogen was successfully produced from mixed xylose/arabinose. The optimum conditions for hydrogen production were the initial concentration of mixed xylose/arabinose 5 g/L each, initial cultivation pH 5.5 and temperature 55 °C. Under the optimum conditions, a maximum hydrogen yield of 2.49 mol-H₂/mol-sugar consumed was obtained. The optimum conditions were then used to produce hydrogen from sugar derived from acid-hydrolysed sugarcane bagasse (SCB) at a reducing sugar concentration of 10 g/L in which a lower hydrogen yield of 1.48 mol-H₂/mol-sugar consumed was achieved. Main soluble product was acetate suggesting the hydrogen fermentation from mixed xylose/arabinose is the acetate type. The dominant hydrogen producers found in both fermentation broth were Thermoanaerobacterium thermosaccharolyticum and Clostridium sp. Lower hydrogen yield in the SCB hydrolysate fermentation broth may be due to the present of Clostridium ragsdalei and microorganisms in the class Bacilli viz. Lactococus lactis subsp., Lactobacillus delbrueckii, and Sporolactobacillus sp. as well as the inhibitors (acetic acid and furfural) contained in the SCB hydrolysate.     

Enhancement of fermentative hydrogen/ethanol production from cellulose using mixed anaerobic cultures

Batch tests were conducted to evaluate the enhancement of hydrogen/ethanol (EtOH) productivity using cow dung microflora to ferment α-cellulose and saccharification products (glucose and xylose). Hydrogen/ethanol production was evaluated based on hydrogen/ethanol yields (HY/EY) under 55 °C at various initial pH conditions (5.5–9.0). Our test results indicate that cow dung sludge is a good mixed natural-microflora seed source for producing biohydrogen/ethanol from cellulose and xylose. The heat-pretreatment, commonly used to produce hydrogen more efficiently from hexose, applied to mixed anaerobic cultures did not help cow dung culture convert cellulose and xylose into hydrogen/ethanol. Instead of heat-pretreatment, the mixed culture received enrichments cultivated at 55 °C for 4 days. Positive results were observed: hydrogen/ethanol production from fermenting cellulose and xylose was effectively enhanced at increases of 4.8 (ethanol) to 8 (hydrogen) and 2.4 (ethanol) to 15.6 (hydrogen) folds, respectively. In which, the ethanol concentration produced from xylose reached 4–4.4 g/L, an output comparable to that of using heat-treated sewage sludge and better than that (1.25–3 g/L) using pure cultures. Our test results show that for the enriched cultures the initial cultivation pH can affect hydrogen/ethanol production including HY, EY and liquid fermentation product concentration and distribution. These results were also concurred using a denaturing gradient gel electrophoresis analysis saying that both cultivation pH and substrate can affect the enriched cow dung culture microbial communities. The enriched cow dung culture had an optimal initial cultivation pH range of 7.6–8.0 with peak HY/EY values of 2.8 mmol-H₂/g-cellulose, 5.8 mmol-EtOH/g-cellulose, 0.3 mol-H₂/mol-xylose and 1 mol-EtOH/mol-xylose. However, a pH change of 0.5 units from the optimal values reduced hydrogen/ethanol production efficiency by 20%. Strategies based on the experimental results for optimal hydrogen/ethanol production from cellulose and xylose using cow dung microflora are proposed.     

Biohydrogen production from food waste in batch and semi-continuous conditions: Evaluation of a two-phase approach with digestate recirculation for pH control

The research investigated the production of Biohythane in a two-phase anaerobic digestion process treating food waste as substrate. Preliminary batch assays were carried out at initial organic loadings of 15, 20, 25 and 30 kg TVS m⁻³, in stirred 1.5-l reactors at 55 °C. The results showed all hydrogen was produced within the first 24 h after feeding and the highest load tested gave the maximum hydrogen production (0.047 m³ H₂ kg⁻¹VS, H₂ 30%). Similar loadings were then tested in a two-phase system. Hydraulic retention times of 3 and 12 days were applied to the first and second reactor respectively. In order to keep the pH at ∼5.5, either supernatant or whole digestate from the methanogenic reactor was recirculated to the first phase. Results showed that hydrogen was produced (0.117 Nm³ kg⁻¹ VS, 47.7%) when recirculating whole digestate with an organic loading rate of 20 kg TVS m⁻³ day⁻¹.     

Biohydrogen and biogas production from mashed and powdered vegetable residues by an enriched microflora in dark fermentation

This study conducted the utilization of vegetable residues by an enriched microflora inoculum to produce biohydrogen via anaerobic batch reactor. Dark fermentation processes were carried out with 3 kinds of vegetable residue substrates including broccoli (Brassica oleracea var. italica.), onion (Alium cepa Linn.), and sweet potato (Ipomoea batatas (L.) Lam). Vegetable wastes were pretreated into 2 forms, i.e. mashed and powdered vegetable, prior to the fermentation. The substrate used for the biohydrogen production were vegetable residues and inoculum at the vegetable residues/inoculum ratio of 1:1 (based on TS). The digestion processes were performed under 120 rpm speed of shaking bottle in the incubator with control temperature of 35^?C. In this work, the maximum hydrogen production was achieved by anaerobic digestion at mashed onion with bioreactor inoculum that produced total hydrogen of 424.1 mL H₂ with hydrogen yield and hydrogen concentration of 151.67 mL H₂/g VS_added and 43.54%, respectively. In addition, the hydrogen production continues took only 7 days for the vegetables blended with the bioreactor inoculum. Finally, it was found that the high potential of degradation of vegetable wastes an enriched microflora in dark fermentation also showed alternative solution to eliminate agricultural wastes to produce green energy.     

Biohydrogen production from cornstalk wastes by anaerobic fermentation with activated sludge

An anaerobic fermentation process to produce hydrogen from cornstalk wastes was systematically investigated in this work. Batch experiments numbered series I, II and III were designed to investigate the effects of acid pretreatment, enzymatic hydrolysis (enzymatic temperature, enzymatic time and enzymatic pH) on hydrogen production by using the natural sludge as inoculant. A maximum cumulative H₂ yield of 126.22 ml g⁻¹-CS (Cornstalk, or 146.94 ml g⁻¹-TS, Total Solid) and an average H₂ production rate of 9.58 ml g⁻¹-CS h⁻¹ were obtained from fermentation cornstalk with a concentration of 20 g/L and an initial pH of 7.0 at 36 °C through an optimal pretreatment process. The optimal process was that the substrate was soaked with an HCl concentration of 0.6 wt% at 90 °C for 2 h, and subsequently enzymatic hydrolysis for 72 h at 50 °C and pH 4.8 before fermentation. The biogas consisted of only H₂ and CO₂. In addition, the fermentation system was the typical ethanol-type fermentation according to ethanol and acetate as the main liquid by-products.     

Bio hydrogen generation from kitchen waste in an inclined plug flow reactor

The performance of inclined plug-flow reactor of 0.15 m³ for hydrogen generation from kitchen waste using 100 °C heat treated biogas plant slurry as inoculum under solid phase anaerobic digestion was studied. The reactor pH stabilized at 5.6 after 21 days of its operation. 40% VS degradation was obtained and it generated 72 mL H₂/gVS added. The soluble metabolites in the reactor were predominantly butyric acid (45–50%), acetic acid (20–30%) and propionic acid (10–20%).     

Effect of pH on continuous biohydrogen production from liquid swine manure with glucose supplement using an anaerobic sequencing batch reactor

pH is considered as one of the most important factors governing the hydrogen fermentation process. In this project, five pH levels, ranging from 4.4 to 5.6 at 0.3 increments, were tested to evaluate the pH effect on hydrogen production from swine manure supplemented with glucose in an anaerobic sequencing batch reactor system with 16 h of hydraulic retention time (HRT). The optimal hydrogen yield (1.50 mol H₂/mol glucose) was achieved at pH 5.0 when the maximum production rate of 2.25 L/d/L was obtained. Continuous hydrogen production was achieved for over 3 weeks for pH 5.0, 4.7, and 4.4, with no significant methane produced. However, as pH increased to 5.3 and 5.6, methane production was observed in the biogas with concurrent reductions in hydrogen production, indicating that methanogens could become increasingly activated for pH 5.3 or higher. Acetate, propionate, butyrate, valerate, and ethanol were the main aqueous products whose distribution was significantly affected by pH as well.     

Optimizing biohydrogen production from mushroom cultivation waste using anaerobic mixed cultures

The mushroom bag is a polypropylene bag stuffed with wood flour and bacterial nutrients. After being used for growing mushroom for one to two weeks this bag becomes mushroom cultivation waste (MCW). About 150 million bags (80,000 tons) of MCW are produced annually in Taiwan and are usually burned or discarded. The cellulosic materials and nutrients in MCW could be used as the feedstock and nutrients for anaerobic biohydrogen fermentation. This study aims to select the inoculum from various waste sludges (sewage sludge I, sewage sludge II, cow dung and pig slurry) with or without adding any extra nutrients. A batch test was operated at a MCW concentration of 20 g COD/L, temperature 55 °C and an initial cultivation pH of 8. The results show that extra nutrient addition inhibited hydrogen production rate (HPR) and hydrogen production yield (HY) when using cow dung and pig slurry seeds. However, nutrient addition enhanced the HPR and HY in case of using sewage sludge inoculum and without inoculum. This related to the inhibition caused by high nutrient concentration (such as nitrogen) in cow dung and pig slurry. Peak HY of 0.73 mmol H₂/g TVS was obtained with no inoculum and nutrient addition. However, peak HPR and specific hydrogen production rate (SHPR) of 10.11 mmol H₂/L/d and 2.02 mmol H₂/g VSS/d, respectively, were obtained by using cow dung inoculum without any extra nutrient addition.